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Updated: Dec 10, 2025
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Preparation of rAAV9 to Overexpress or Knockdown Genes in Mouse Hearts
Published on: December 17, 2016
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AAV Gene Transfer to the Heart
Suya Wang1, Yuxuan Guo1, William T Pu2
1Department of Cardiology, Boston Children's Hospital, Boston, MA, USA.
Methods in Molecular Biology (Clifton, N.J.)
|August 29, 2020
Summary
This study details a recombinant adeno-associated virus (rAAV) production method for gene therapy. The protocol yields significant viral particles, demonstrating rAAV9
Area of Science:
- Gene therapy
- Molecular biology
- Virology
Background:
- Recombinant adeno-associated virus (rAAV) is crucial for gene therapy, enabling sustained protein expression in targeted tissues.
- Effective gene delivery relies on efficient viral vector production and purification.
Purpose of the Study:
- To describe a robust protocol for producing high-titer rAAV vectors.
- To evaluate the efficacy of rAAV9 for cardiac gene delivery in vivo.
Main Methods:
- PEI-mediated triple transfection of HEK293T cells for rAAV production.
- Purification of rAAV vectors using iodixanol density gradient ultracentrifugation.
Main Results:
- The described protocol typically yields 3E11 viral genomes (vg) per production run.
- rAAV9 demonstrated significant transduction of cardiac cells following in vivo injection.
Conclusions:
- The developed rAAV production protocol is efficient and scalable.
- rAAV9 is a promising vector for cardiac gene therapy applications.
